Systems and methods of delivery of bioactive agents using bacterial toxin-derived transport sequences

Abstract

The field of the present invention relates, in part, to a strategy for novel pharmaceutical applications. More specifically, the present invention relates to a non-toxic mutant form of the Vibrio cholera Cholix gene (ntCholix), a variant of Cholix truncated at amino acid A 386 (Cholix 386 ) and the use of other various Cholix-derived polypeptide sequences to enhance intestinal delivery of biologically-active therapeutics. Importantly, the systems and methods described herein provide for the following: the ability to deliver macromolecule doses without injections; the ability to deliver cargo, such as (but not limited to) siRNA or antisense molecules into intracellular compartments where their activity is required; and the delivery of nanoparticles and dendrimer-based carriers across biological membranes, which otherwise would have been impeded due to the barrier properties of most such membranes.

What is claimed is: 
     
         1 . An isolated delivery construct comprising:
 (a) a non-toxic Cholix toxin, wherein the Cholix construct has a mutation in domain Ill that renders the construct non-toxic; and   (b) a therapeutic cargo.   
     
     
         2 . The isolated delivery construct according to  claim 1 , wherein the delivering is oral. 
     
     
         3 . The isolated delivery construct according to  claim 1 , further comprising one or more cleavable linker. 
     
     
         4 . The isolated delivery construct according to  claim 1 , wherein the therapeutic cargo is a macromolecule or small molecule selected from the group consisting of siRNA, PNA, miRNA, DNA, plasmid and antisense molecules. 
     
     
         5 . A pharmaceutical composition comprising
 (a) an isolated delivery construct according to  claim 1 ; and   (b) a pharmaceutically acceptable carrier.   
     
     
         6 . The pharmaceutical composition according to  claim 5 , wherein the delivery construct is formulated for oral administration, topical administration, pulmonary administration, intra-nasal administration, buccal administration, sublingual administration or ocular administration. 
     
     
         7 . The pharmaceutical composition according to  claim 5 , wherein the delivery construct is formulated for oral administration. 
     
     
         8 . The pharmaceutical composition according to  claim 5 , wherein the composition is formulated in a capsule or tablet. 
     
     
         9 . The pharmaceutical composition according to  claim 5 , wherein the composition is formulated for oral administration. 
     
     
         10 . The pharmaceutical composition according to  claim 5 , comprising from about 1 μg to about 1 g of the isolated delivery construct. 
     
     
         11 . The pharmaceutical composition according to  claim 5 , wherein the isolated delivery construct is from about 10 μg to about 100 mg. 
     
     
         12 . The pharmaceutical composition according to  claim 5 , wherein the isolated delivery construct is from about 10 μg to about 1000 μg.